A COMPARISON OF THE SENSITIVITY OF IMMUNOPEROXIDASE STAINING METHODS WITH HIGH-SENSITIVITY FLUORESCENCE FLOW-CYTOMETRY ANTIBODY QUANTITATION ON THE CELL-SURFACE
A COMPARISON OF THE SENSITIVITY OF IMMUNOPEROXIDASE STAINING METHODS WITH HIGH-SENSITIVITY FLUORESCENCE FLOW-CYTOMETRY ANTIBODY QUANTITATION ON THE CELL-SURFACE
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DOI：
10.1177/42.8.8027533
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发表时间：
1994-08-01
影响因子：
3.2
通讯作者：
BRADLEY, J
中科院分区：
文献类型：
--
作者：
COVENTRY, BJ;NEOH, SH;BRADLEY, J
以低拷贝数存在的表面分子可以用高灵敏度荧光流式细胞术检测。许多以前被认为在其表面不表达某些分子的细胞现在可以通过高灵敏度荧光细胞计数方法显示这些分子的拷贝数非常低。通过免疫过氧化物酶染色方法检测分子以前没有与高灵敏度流式细胞术技术进行比较。计算机化视频图像分析（VIA）是一种允许以类似于流式细胞术的标准化方式测量免疫染色显色反应产物的面积和密度的方法。在这项研究中，我们比较了免疫过氧化物酶反应产物通过VIA方法测量与高灵敏度现在细胞计数测量与10,000到50个抗体分子结合到其表面的细胞。100-200个表面分子的检测是可能的重金属增强免疫过氧化物酶法，而标准的免疫过氧化物酶法不敏感。镍增强的免疫过氧化物酶染色方法的灵敏度被证实用于检测外周血淋巴细胞表面上以低数量存在的表位（Tac-IL 2受体α链）。
Surface molecules present in low copy numbers can be detected with high-sensitivity fluorescence now cytometry. Many cells previously thought not to express certain molecules on their surface can now be shown to have these molecules in very low copy numbers by high-sensitivity fluorescent cytometric methods. Detection of molecules by immunoperoxidase staining methods has not previously been compared with high-sensitivity flow cytometry techniques. Computerized video image analysis (VIA) is a method that allows measurement of area and density of the immunostain chromogen reaction product in a standardized fashion analogous to flow cytometry. In this study, we compared immunoperoxidase reaction products measured by VIA methods with high-sensitivity now cytometric measurements for cells with 10,000 down to 50 antibody molecules bound to their surfaces. Detection of 100-200 surface molecules was possible with heavy metal-enhanced immunoperoxidase methods, whereas standard immunoperoxidase methods were not as sensitive. The sensitivity of the nickel-enhanced immunoperoxidase staining method was confirmed for detection of an epitope (Tac-IL2 receptor alpha-chain) present in low numbers on the surface of peripheral blood lymphocytes.